A simplified microassay for serotonin: Modification of the enzymatic isotopic assay |
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Authors: | Ilan Hammel Yehudith Naot Ester Ben-David Haim Ginsburg |
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Affiliation: | Department of Immunology Faculty of Medicine Technion-Israel Institute of Technology Haifa, Israel |
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Abstract: | A simplification of the enzymatic isotopic assay for serotonin is described, Serotonin is converted to [3H]melatonin by a two-step reaction: N-acetylation of serotonin using acetic anhydride, followed by O-methylation with the enzyme hydroxyindole O-methyltransferase (EC 2.1,1.4) and S-adenosyl--[methyl-3H]methionine as methyl donor. The present assay avoids the use of unstable acetylating enzyme, rat liver N-acetyltransferase (EC2.3,1.5). Blank values are lowered considerably and the sensitivity is doubly increased. Two-tenths micromole of serotonin per 30 μl of sample in tissue homogenates can be measured. |
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